Note: Descriptions are shown in the official language in which they were submitted.
CA 02862545 2014-07-24
BIODEGRADABLE NON-WOVEN MATERIAL FOR MEDICAL PURPOSES
The invention relates to a biodegradable fleece, a method for producing said
biodegradable
fleece as well as the use of said biodegradable fleece as local haemostatic
agent.
Surgical interventions are often associated with local haemorrhage in soft
tissues that cannot be
staunched with common methods of haemostasis, such as direct compression,
suturing, clips or
cauterisation. Effective haemostasis during surgical interventions can clearly
reduce the number
of transfusions needed and improve the visualisation of the site of
intervention and reduce the
surgery time. Moreover, effective haemostasis also reduces the mortality and
morbidity of the
patients during and after surgical interventions. For this reason, sponges,
films, gauze materials
and powders made of collagen, cellulose and/or gelatine have been developed
for use as local
passive haemostatic agents.
It is a disadvantage, especially of the powders, that these often adhere to
gloves and instruments
of the physicians performing the surgery as a result of electrostatic effects,
which makes them
difficult to handle. The haemostatic effect of said sponges or films or gauze
materials, is based
on platelet aggregation at the surface of the proteins or cellulose, of which
these are made. This
enables the formation of a thrombus and effective closure of the defect.
Regarding collagen
haemostatic agents, it needs to be taken into account that between 2% and 4%
of the total popu-
lation are allergic to bovine collagen [A. K. Lynn, I. V. Yannas, W. Bonfield;
Antigenicity and
immunogenicity of collagen; J Biomed Mater Res B Appl Biomater. 2004; 71(2);
343-354]. Cel-
lulose-based products contain regenerated oxidised cellulose. There is some
evidence in the lit-
erature indicating that these are absorbed less well than collagen and
gelatine-based products.
Several case studies have shown that it was possible to identify residues of
the oxidised cellulose
at a revision surgery [Y. Tomizawa; Clinical benefits and risk analysis of
topical hemostats: a
review; J. Artif. Organs. 2005; 8(3); 137-142]. Due to its properties,
gelatine can also be used in
the case of irregularly shaped wound geometries. Affixing a haemostatic agent
of this type to the
site of haemorrhage, the material adapts to the wound and swells which
produces a tamponade
effect in confined spaces. Swollen gelatine reduces the blood flow and forms a
stable matrix
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CA 02862545 2014-07-24
about which a thrombus is formed by means of contact activation. Essentially,
there are gelatine-
based products of bovine and porcine origin available.
A disadvantage in terms of the handling during surgical interventions is the
high tackiness of the
blood-soaked products on surgical instruments [S. Srinath; Topical hemostatic
agents in surgery:
A surgeon's perspective; Aorn Journal. 2008; 88(3) 2-11]. Sponges are
generally produced by
freeze drying and also by special foaming processes. This is disadvantageous
since fibroblasts
can migrate only with difficulty or not at all into the sponges and swollen
powders in the scope
of wound healing. In general, the use of excessive quantities of such solely
passive haemostatic
agents that are based on collagen, gelatine, and in particular cellulose, has
been observed to be
associated with complications. Residues of the product can cause foreign body
reactions, chronic
inflammations and/or infections at the site of use, which, in turn, promote
the formation of
granuloma and prevent optimal healing. Granuloma has been observed at a wide
variety of sites
with solely passive haemostatic agents [H. E. Achneck, B. Sileshi, R. M.
Jamiolkowski, D. A.
Albala, M. L. Shapiro, J. H. Lawson; A comprehensive review of topical
hemostatic agents: Ef-
ficacy and recommendations for use; Annals of Surgery. 2010; 251(2). 217-228].
The process of coagulation is sub-divided into primary haemostasis and
secondary haemostasis.
The essential step of primary haemostasis is platelet aggregation, which leads
to initial closure of
the bleeding. Secondary haemostasis is a complex cascading process at the end
of which fibrin is
released from fibrinogen by the thrombin protease and forms a stable fibrin
network through
cross-linking. Secondary haemostasis can be triggered, inter alia, by calcium
ions, i.e. factor IV.
A number of active haemostatic agents based on collagen sponges containing
thrombin has been
proposed for activating the formation of fibrin from fibrinogen upon contact
to blood. Said active
haemostatic agents show biological activity and intervene directly in the
later phases of the com-
plex cascading process in order to induce a thrombus at the site of
haemorrhage. This is to
staunch the bleeding rapidly. The presence of fibrinogen in the patient's
blood is required for
effective haemostasis by means of thrombin, which therefore fails in
afibrinogemaemia patients.
However, it must be viewed critically, especially regarding the use of human
thrombin, that the
thrombin needs to be treated appropriately such that any transmission of
viruses, such as HIV
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CA 02862545 2014-07-24
and HCV, can be safely excluded. Moreover, bovine and human thrombins have
been observed
to possess a potential to induce antibodies (in up to 94% of the cases) [H.
Seyedejad, M. Imani,
T. Jamieson, A. M. Seifalian; Topical haemostatic agents; British Journal of
Surgery; 2008; 95;
1197-1225]. Although many patients show no clinical anomalies after the
development of anti-
bodies, anomalies have indeed been observed in blood coagulation tests, even
with fatal out-
comes in exceptional cases (anaphylaxis, coagulopathy) [Y. Wai, V. Tsui, Z.
Peng, R. Richard-
son, D. Oreopoulos, S. M. Tarlo; Anaphylaxis from topical bovine thrombin
during haemodialy-
sis and evaluation of sensitization among dialysis population; Clin Exp
Allergy; 2003; 33; 1730-
1734; M. Pope, K. W. Johnston; Anaphylaxis after thrombin injection of a
femoral pseudoaneu-
rysm: recommendations for prevention; J Vasc Surg; 2000; 32; 190-191; und K.
Tadokoro,
T.Ohtoshi, S. Takafuiji, K. Nakajima, S. Suzuki, K. Yamamoto et al.; Topical
thrombin-induced
IgE-mediated anaphylaxis: RAST analysis and skin-test studies; .1 Allergy Clin
Immunol 1991;
88; 620-629]. In addition, the use of bovine thrombin in the human body has
been observed to be
associated with severe immune defence reactions.
In the human body, the plasmin protease acts as antagonist of secondary
haemostasis. Plasmin
cleaves fibrin networks into small fragments. This process called fibrinolysis
counteracts secon-
dary haemostasis.
The haemostatic fleeces thus known are disadvantageous in that haemostasis is
not attained rap-
idly and effectively enough in some cases. Specifically, it is a disadvantage
of the haemstatic
fleeces according to the prior art that the effect of secondary haemostasis is
reduced by the
body's inherent plasmin protease and that the haemostatic effect of the
haemostatic fleeces is thus
limited.
It is therefore the object of the invention to provide an improved
biodegradable fleece which can
preferably be used to overcome the afore-mentioned disadvantages. In
particular and preferably,
a biodegradable fleece is to be provided that possesses a stronger haemostatic
effect than previ-
ous haemostatic fleeces. Concurrently, the biodegradable fleece should be easy
to use and as
inexpensive as possible to manufacture.
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CA 02862545 2016-01-08
Specifically, a fleece is to be developed that activates both primary and
secondary haemostasis
and in which the fibrin network thus produced is further stabilised. Moreover,
the nature of the
fleece should be appropriate such that hum in fibroblasts can migrate into the
fleece such that
connective tissue can be formed in the cour ;e of wound healing. The fleece
should, if possible,
not contain proteins isolated from human blood in order to sidestep any
transmission of infective
pathogens, in particular of human viruses.
It is another goal to preferably design the fleece appropriately such that the
pH value of the
fleece is being stabilised in the physiological neutral pH range such that
wound healing cannot
be impaired by any shifts in pH.
Moreover, it should be feasible to modify ihe fleece with anti-infective
agents such that local
protection of the fleece against microbial cohnisation can be attained.
Accordingly, the invention provides a biodegradable fleece containing (i) at
least one polymer
for inducing primary haemostasis, (ii) at 1,..ast one non-proteinogenic, low-
molecular weight,
water-soluble activator of secondary haemo3tasis, and (iii) at least one non-
proteinogenic, low-
molecular, water-soluble inhibitor of fibrinolysis.
Moreover, the invention provides a method fòr producing said biodegradable
fleece, whereby (i)
a fluidised fibre raw material, and additives if applicable, is placed in a
container, (ii) the con-
tainer is made to rotate, (iii) the fluidised fi Dre raw material is dispensed
from the container by
means of centrifugal forces, whereby fibres 1) or filaments (1) are formed,
and (iv) a biodegrad-
able fleece is produced from the fibres (1) or filaments (1).
Moreover, the invention provides the use of said biodegradable fleece as local
haemostatic agent.
Presently, a biodegradable material shall be understood to mean materials, in
particular poly-
mers, and components that degrade and are absorbed under in-vivo conditions.
The materials are
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CA 02862545 2016-01-08
eliminated via the natural metabolic pathway in this context. This can involve
simple filtration
processes of the degradation products or proceed after their metabolisation.
The biodegradable fleece contains (i) a polymer for inducing primary
haemostasis.
The polymer for inducing primary haemostasis is preferably selected from the
group consisting
of collagen, gelatine, carboxymethylcellulose, oxycellulose,
carboxymethyldextran, and mixtures
thereof. These polymers are readily available and are particularly well-suited
for building-up the
haemostatic fleece and/or the fibres for the haemostatic fleece. The fleece
contains at least one
non-proteinogenic, low-molecular, water-soluble activator of secondary
haemostasis.
The activator of haemostasis is non-proteinogen, if it comprises no a-amino
acids, no peptides
and no oligopeptides, preferably no peptides at all.
The activator is low-molecular weight, if its molar mass is less than 1,000
g/mol. Preferably, the
activator has a molar mass of less than 800 g/mol, more preferably of less
than 500 g/mol, and
particularly preferably of less than 200 g/mol.
Preferably, the activator of secondary haemostasis is soluble in water if the
solubility of the acti-
vator of secondary haemostasis in water at a temperature of 25 C is at least
100 mg per litre,
more preferably at least 500 mg per litre, even more preferably at least 1,000
mg per litre, and
particularly preferably at least 2,000 mg per litre.
Said activator of secondary haemostasis preferably acts haemostatic, i.e. it
is well-suited to coun-
teract by medical means a bleeding in a patient. Preferably, the activator of
secondary haemosta-
sis supports the inherent haemostasis of the body such that the bleeding is
staunched more rap-
idly.
According to a preferred embodiment, the activator of secondary haemostasis is
at least one cal-
cium salt. Said at least one calcium salt preferably has a solubility in water
of more than 2 g/litre
at a temperature of 25 C. Preferably, the at least one calcium salt is
selected from the group con-
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CA 02862545 2016-01-08
sisting of calcium chloride, calcium acetate, calcium sulfate dihydrate,
calcium lactate, and mix-
tures thereof. Calcium salts can be used particularly easily in the build-up
of a fleece according
to the invention. Moreover, they can be converted easily by the organism of
the patient.
The fraction of the activator of secondary haemostasis preferably is in the
range of 0.1 to 20 %
by weight, more preferably in the range of 0.5 to 15 % by weight, and even
more preferably in
the range of 1 to 10 % by weight, relative to the weight of the fleece.
The fleece contains at least one non-proteinogenic, low-molecular weight,
water-soluble inhibi-
tor of fibrinolysis.
The inhibitor of fibrinolysis is non-proteinogen, if it comprises no a-amino
acids, no peptides
and no oligopeptides, preferably no peptides at all.
The inhibitor of fibrinolysis is low-molecular weight, if its molar mass is
less than 1,000 g/mol.
Preferably, the inhibitor of fibrinolysis has a molar mass of less than 800
g/mol, more preferably
of less than 500 g/mol, and particularly preferably of less than 200 g/mol.
Preferably, the inhibitor of fibrinolysis is soluble in water if the
solubility of the inhibitor of fi-
brinolysis in water at a temperature of 25 C preferably is at least 100 mg per
litre, more prefera-
bly at least 500 mg per litre, even more preferably at least 1,000 mg per
litre, and particularly
preferably at least 2,000 mg per litre.
According to a preferred embodiment of the invention, the non-proteinogenic,
low-
molecular weight, water soluble inhibitor of fibrinolysis is a lysine
analogue. Prefera-
bly, the non-proteinogenic, low-molecular weight, water-soluble inhibitor of
fibri-
nolysis is an amphoteric aminocarboxylic acid. Preferably, the non-
proteinogenic,
low-molecular weight, water-soluble inhibitor of fibrinolysis is an a-
aminocarboxylic
acid. Preferably, the at least one non-proteinogenic, low-molecular weight,
water-
soluble inhibitor of fibrinolysis is selected from the group consisting of 6-
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CA 02862545 2016-01-08
aminohexanoic acid, 4-aminomethylbenzoic acid, trans-4-
aminomethylcyclohexylcarboxylic
acid and mixtures thereof. Said substances have proven to be particularly well-
suited for produc-
ing the haemostatic fleece and/or for producing the fibres for the haemostatic
fleece. Moreover,
said substances are non-objectionable for the patient from a medical point of
view and can there-
fore be used in medical devices.
The invention can just as well provide that the amount of the non-
proteinogenic, low-molecular
weight, water-soluble inhibitor of fibrinolysis is selected appropriately such
same has a pH-
stabilising buffering effect at the surface of the biodegradable fleece,
whereby the inhibitor pref-
erably buffers the pH value in the range between 6 and 8.
The inhibitor of fibrinolysis having a buffering effect is particularly
advantageous because there
is then no need to introduce an additional substance as buffer into the
biodegradable fleece.
Preferably the fraction of the inhibitor of fibrinolysis is in the range of
0.1 to 20 % by weight,
more preferably in the range of 0.5 to 15 % by weight, and even more
preferably in the range of
1 to 10 % by weight, relative to the weight of the fleece. With the fractions
of the inhibitor of
fibrinolysis being as indicated, fibrinolysis is inhibited to a sufficient
degree and the pH value is
maintained in a preferred range for haemostasis.
According to a preferred embodiment, the fibres of the biodegradable fleece
comprise (i) the
polymer for inducing primary haemostasis, (ii) the non-proteinogenic, low-
molecular weight,
water-soluble activator of secondary haemostasis, and/or (iii) the non-
proteinogenic, low-
molecular weight, water-soluble inhibitor of fibrinolysis. It is preferable in
this context that (i)
the polymer for inducing primary haemostasis, (ii) the non-proteinogenic, low-
molecular weight,
water-soluble activator of secondary haemostasis, and/or (iii) the non-
proteinogenic, low-
molecular weight, water-soluble inhibitor of fibrinolysis are distributed
homogeneously in the
fibres of the fleece.
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CA 02862545 2014-07-24
Moreover, the invention can provide the haemostatic fleece to comprise at
least one anti-
infective agent.
Preferably, said at least one anti-infective agent is an antibiotic.
Preferably, said at least one anti-infective agent is present in the fibres of
the biodegradable
fleece. In this context, said at least one anti-infective agent can just as
well be arranged on the
fleece surface.
Preferably, said at least one anti-infective agent is soluble in water.
Preferably, the at least one
anti-infective agent is soluble in water if the solubility of the at least one
anti-infective agent in
water at a temperature of 25 C preferably is at least 100 mg per litre, more
preferably at least
500 mg per litre, even more preferably at least 1,000 mg per litre, and
particularly preferably at
least 2,000 mg per litre.
Preferably, a pharmaceutically effective amount of the at least one anti-
infective agent is present
in the fleece.
It is of particular advantage that the at least one anti-infective agent is co-
incorporated into the
fleece during the production of the biodegradable fleece. For example, the at
least one anti-
infective agent can be taken up into the fibre material for producing the
biodegradable fleece in
this context. Said fleeces possess an additional anti-inflammatory effect and
counteract an infec-
tion of the patient.
The invention can just as well provide the biodegradable fleece to comprise a
buffer substance
that is poorly soluble in water. Said buffer substance can be present, for
example, in the fibres of
the fleece and can be distributed homogeneously in the fibres of the fleece,
if applicable.
Preferably, a buffer substance is poorly soluble in water if the solubility of
the buffer substance
in water at a temperature of 25 C is less than 10 g and more preferably less
than 5 g. The solubil-
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CA 02862545 2014-07-24
ity of the buffer substance in water at a temperature of 25 C preferably is in
the range of
1 mg/litre to 1 g/litre and more preferably in the range of 5 mg/litre to 500
mg/litre.
According to a preferred embodiment, the buffer substance is selected from the
group consisting
of calcium carbonate, magnesium carbonate, basic magnesium carbonate, and
mixtures thereof.
An additional buffer substance helps in adjusting a suitable pH value for
haemostasis in the
blood of the patient in the immediate vicinity of the biodegradable fleece
inserted into the
wound.
Another refinement of the invention can provide the biodegradable fleece to
comprise a pH indi-
cator. In this context, the fibres of the fleece, in particular, can comprise
a pH indicator.
Said pH indicator preferably has a transition point at a pH of less than pH
7.4. Preferably, the pH
indicator is bromocresol purple or bromothymol blue.
The pH indicator can be used to visually check the situation existing at the
wound site or whether
further treatment measures are required in order to achieve rapid haemostasis.
According to a particularly preferred refinement of the invention, the average
mesh width be-
tween the fibres of the dry fleece is at least 5011111. The average mesh width
between the fibres of
the dry fleece preferably is in the range of 50 um to 500 um and more
preferably in the range of
100 l.tm to 200 um.
A mesh width being within the range specified above allows fibroblasts to grow
into the fleece.
This effects more rapid wound healing upon the use of a biodegradable fleece
according to the
invention.
The invention can just as well provide the fibres of the biodegradable fleece
to comprise a mean
fibre diameter in the range of 0.5 um to 500 um, preferably in the range of 2
um to 300 um, and
more preferably in the range of 5 um to 200 um.
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Fibres of the diameter specified above are sufficiently strong to prevent
individual fibres from
breaking and to provide sufficient surface area to the biodegradable fleece in
order to dispense
sufficient concentrations of the agents.
The biodegradable fleece according to the invention can be used, for example,
as local haemo-
static agent.
The invention is based, in part, on the surprising finding that combining an
activator of secon-
dary haemostasis and an inhibitor of fibrinolysis as water-soluble components
of a biodegradable
fleece improves the haemostatic effect.
As mentioned above, the human organism has the plasmin protease as an
antagonist of secondary
haemostasis that cleaves the fibrin network into small fragments.
The process of fibrinolysis therefore counteracts secondary haemostasis. The
formation of plas-
min is induced by plasminogen activators.
Plasmin can be inhibited by analogues of the amino acid, lysine (K. Aktories,
U. Forstermann,
W. Forth; Allgemeine und spezielle Pharmakologie und Toxikologie; Elsevier,
Urban&Fischer
Verlag, 9. edition, 2006; 547). Known for this purpose are, inter alia, a-
aminocarboxylic acids,
whereby the amino group and the carboxyl group have to be separated by five
carbon atoms.
Commercially available haemostatic preparations containing the anti-
fibrinolytic agent,
aprotinin, are associated with significant disadvantages. Bovine aprotinin can
trigger anaphylac-
tic reactions [R. N. Kaddoum, E. J. Chidiac, M. M. Zestos, S. D. Rajan, A.
Baraka; An anaphy-
lactic reaction after primary exposure to an aprotinin test dose in a child
with a severe milk al-
lergy; J. Cardiothorac. Vasc. Anesth.; 2007; 21; 243-244]. Between 0.9% and
2.6% of the pa-
tients treated with bovine aprotinin showed hypersensitivity reactions upon
repeated exposure
[W. Dietrich, A. Ebell, R. Busley, A. Boulesteix; Aprotinin and anaphylaxis:
analysis of 12403
exposures to aprotinin in cardiac surgery; Ann. Thorac. Surg.; 2007; 84; 1144-
1150].
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CA 02862545 2014-07-24
It has been been found, surprising,ly, in the scope of the invention that
inhibitors of fibrinolysis,
such as lysine analogues, can be used as water-soluble (and thus blood-
soluble) components of
haemostatic fleeces in order to improve the haemostatic effect of the fleece.
The invention is also, in part, based on a surprising effect, i.e. that lysine
analogues, as low-
molecular inhibitors of fibrinolysis, incorporated in sufficient amounts into
the fleece, buffer the
pH value of the surface and immediate vicinity of the fleece to a
physiological, nearly neutral pH
range.
The prior art includes no local haemostatic material in the form of sponges,
wovens or fleeces,
which induce primary and secondary haemostasis and concurrently inhibit the
fibrinolysis of the
fibrin network thus produced.
The invention provides a biodegradable fleece that activates both primary and
secondary haemo-
stasis and in which the fibrin network thus produced is further stabilised
through inhibition of
fibrinolysis.
Moreover, the prior art also includes no local haemostatic material that
adjusts the pH value in
the immediate vicinity of the haemostatic material in a targeted manner
through addition of a
neutral range buffer and thus promotes haemostasis through this measure as
well.
The pH value of the wound exudate influences the wound healing process. For
optimal wound
healing, a neutral pH vaue is of advantage (J. Dissemond; Die Bedeutung des pH-
Wertes flir die
Wundheilung; HARTMANN wundForum 1/2006; 15-19). Therefore, adjusting the pH
value to
within a desired range according to the invention is also advantageous in the
use of fleeces ac-
cording to the invention. For this purpose, a soluble buffer substance is
applied to the fleece. It is
particularly advantageous and attains a special additive effect to use the
inhibitor of fibrinolysis,
which is used anyway, also as buffer substance. For this purpose, it is
necessary to simply apply
a sufficient amount of the inhibitor to the surface of the fleece such that it
has a pH-stabilising
buffering effect in the immediate vicinity of the surface of the fleece.
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The biodegradable fleece according to the invention can preferably be produced
by means of the
method for producing a biodegradable fleece as described herein.
In said method (i) a fluidised fibre raw material, and additives if
applicable, is placed in a con-
tainer, (ii) the container is made to rotate, (iii) the fluidised fibre raw
material is dispensed from
the container by means of centrifugal forces, whereby fibres (1) or filaments
(1) are formed, and
(iv) a biodegradable fleece is produced from the fibres (1) or filaments (1).
Preferably, the con-
tainer, in which the fluidised fibre material, and additives if applicable,
are placed is a spinning
rotor.
This production method is particularly easy and inexpensive to implement.
In this context, the invention can provide the fibres and filaments thus
produced to be captured as
a two-dimensional material upon their exit from the rotating container,
whereby connecting sites
between two or more fibres are generated in a multitude of regions of the two-
dimensional mate-
rial.
This measure also serves to keep the production method simple and inexpensive.
Moreover, the invention can provide the fleece, in particular the two-
dimensional material, to be
soaked and/or coated with at least one fluid medium in at least one post-
treatment step, whereby,
in particular, biologically degradable polymer materials and/or wax-like
materials are used as
liquid medium.
Biodegradable fleeces according to the invention can be produced in simple and
inexpensive
manner using a rotation spinning method, for example according to DE 10 2007
011 606 A 1,
WO 2011/116848, and DE 10 2007 044 648 A1.
The fibres and/or filaments produced by the spinning rotor are easy to capture
in a condition, in
which connecting sites between two or more fibres are generated in a multitude
of regions of the
two-dimensional material.
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CA 02862545 2014-07-24
In an optional post-treatment step, a large number of properties of the two-
dimensional material
according to the invention can be adapted to specific applications.
By cross-linking the material, the mechanical and, in particular, the chemical
properties of the
biodegradable fleece can be modified. For example, the absorption properties
for medical appli-
cations can be defined through the degree of cross-linking of the material.
The two-dimensional materials according to the invention can be soaked and/or
coated with liq-
uid media in post-treatment steps. For this purpose, in particular but not
exclusively, other bio-
logically degradable polymer materials or wax-like materials are conceivable.
The method according to the invention described above can be used to easily
produce two-
dimensional materials of fibres for biodegradable fleeces according to the
invention whose fibres
have a mean fibre thickness of 0.5 !am to 500 lam.
For producing partially cross-linked materials in the fibres, it is preferred
to add a cross-linker
already to the spinning solution. However, the already spun fibres can be
cross-linked also and
additionally by contacting them to a cross-linker, either in gaseous form or
in solution.
According to the invention, finished randomly-oriented mats can be subjected
to further cross-
linking, which then determines the final degree of cross-linking of the fibres
in the two-
dimensional material and thus the biological degradation rate thereof.
Various methods are available for cross-linking, whereby enzymatic methods,
the use of com-
plexing agents or chemical methods are preferred.
In chemical cross-linking, the cross-linking is performed by means of one or
more reactants, in
particular using aldehydes selected from formaldehyde and dialdehydes,
isocyanates, diisocy-
anates, carbodiimides, alkyldihalogenides. Moreover, hydrophilic di- and
trioxiranes such as, for
example, 1,4-butanediol diglycidyl ether, glycerol triglycidyl ether, and
polyethylene glycol de-
rivatives can be used. The use of polyethylene glycol diglycidylether is
particularly preferred in
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CA 02862545 2016-01-08
anates, carbodiimides, alkyldihalogenides. Moreover, hydrophilic di- and
trioxiranes such as, for
example, 1,4-butanediol diglycidyl ether, glycerol triglycidyl ether, and
polyethylene glycol de-
rivatives can be used. The use of polyethylene glycol diglycidylether is
particularly preferred in
this context. Aside from cross-linking, polyethylene glycol derivatives showed
the beneficial
property to prevent undesired adhesions, for example pericardial adhesions in
the case of heart
surgeries [W. F. Konertz, M. Kostelka, F. W. Mohr et al.; Reducing the
incidence and severity of
pericardial adhesions with a sprayable polymeric matrix; Ann. Thorac. Surg.;
2003; 76; 1270-
1274].
In particular with regard to medical applications, it is recommended to remove
any excess of the
cross-linker from the two-dimensional material and/or the randomly-oriented
mat after cross-
linking.
As described above, it is preferred to add a cross-linker already to the
spinning solution and to
then perform a further cross-linking on the finished two-dimensional material,
basically in a sec-
ond stage, up to the desired degree of cross-linking.
The invention can provide the porosity c of a biodegradable fleece to be given
or calculated by
the following formula:
_ PV fleece
PBulk MVfleece
1 VV fleece
=
ma in, mc
Pa(ma + ma + mc) + (Ma + mb + Mc) 4- Pc = (ma + mb + mc)
Here, p fleece is the density of the non-compressed biodegradable fleece, P
Bulk is the density of the
fibres of the biodegradable fleece, Mfleece is the mass of the biodegradable
fleece, Vfleece is the vol-
ume of the biodegradable fleece, pa is the density of the fibre-forming
polymer, pb is the density
of the activator of secondary haemostasis, pc is the density of the inhibitor
of flbrinolysis, ma is
the mass of the fibre-forming polymer in the fleece, mb is the mass of the
activator of secondary
haemostasis in the fleece, and mc is the mass of the inhibitor of fibrinolysis
in the fleece.
If further components are present in the biodegradable fleece, such as, for
example, an additional
buffer substance or antibiotics, and the porosity of the biodegradable fleece
is to be determined,
14
CA 02862545 2016-01-08
further parameters need to be taken into consideration according to the same
pattern as shown
above, i.e. the masses (md, me, ...) and densities (Pa, Pe, ...) of the
additional components.
It is particularly preferred according to the invention that the fibre surface
Ofibõ calculated or
given according to the following formula:
1 ¨ E = V fleece
fibre = ' fibre = 2
(0fibre)
2
whereby Ofibre is the average diameter of the fibres.
Exemplary embodiments of the invention shall be illustrated in the following
on the basis of four
schematic figures, though without limiting the scope of the invention. The
terms, fleece and non-
wovens, are used as synonyms in the exemplary embodiments. In the figures:
Figure 1: shows a gelatine fleece according to the invention with dry anti-
microbial substance;
Figure 2: shows a gelatine fleece according to the invention with wet anti-
microbial substance in
water;
Figure 3: shows a gelatine fleece according to the invention with anti-
microbial coating; and
Figure 4: shows a gelatine fleece according to the invention with anti-
microbial substance and
anti-microbial coating.
EXEMPLARY EMBODIMENT 1:
Figures 1 and 2 show a first exemplary embodiment of a gelatine fleece
according to the inven-
tion with dry anti-microbial substance (Figure 1) and the gelatine fleece
according to the inven-
tion with wet anti-microbial substance (Figure 2). The fleece comprises fibres
1 or filaments 1
that are situated as a randomly-oriented mat and are cross-linked to each
other. In the wet state
according to Figure 2, the wet fibres 1 are seen to be curved more strongly
than in the dry state
due to the action of the liquid.
15
CA 02862545 2014-07-24
The gelatine fleece with anti-microbial substance according to Figure 1 (dry)
and Figure 2 (wet;
after six hours in distilled water) was produced by means of a rotation
spinning method as fol-
lows:
Firstly, a 24% gelatine solution was prepared. It is conceivable to use a
gelatine of type A PIG-
SKIN of GELITA AG, which was in fact used in the present exemplary embodiment.
The gela-
tine was stirred in water. The pH was adjusted to 7.4 with NaOH (product
number: 3306576,
Sigma-Aldrich, Germany). A total of 1 % by weight calcium chloride (product
number: 102382,
Merck, Germany), 1 % by weight calcium carbonate (product number: C4830, Sigma-
Aldrich,
Germany), 1 % by weight glycerol (product number: 01873, Sigma-Aldrich,
Germany), and
0.5 % by weight 6-aminohexanoic acid (product number: 800145, Merck, Germany)
were added
to the gelatine solution.
The solution was then allowed to stand uninterrupted for approximately one
hour to swell. Then,
the gelatine solution was treated at 60 C in an ultrasonic bath and
maintained at a temperature of
80 C to 85 C for approx. 1 hour. A total of 6 % by weight gentamicin sulfate
(product number:
345814, Merck, Germany) were then dissolved while stirring in the hot gelatine
solution thus
produced.
The gelatine solution maintained at 80 C to 85 C was guided, as the fibre
raw material, by
means of a syringe pump into the container of a device for rotation spinning
according to
DE 10 2005 048 939 Al. A second syringe pump was used concurrently to guide
polyethylene
glycol diglycidylether (product number: 475696, Sigma-Aldrich, Germany) into
the container.
The temperature of the container is approx. 120 C and the container rotates
at a speed of 4,500
rpm. Inside the container there are recesses designed to be holes with a
diameter of 0.3 mm. The
centrifugal force pressed the fibre raw material through said recesses and
spins it into fibres 1
that are drawn by means of an aspiration facility. The aspiration facility was
situated below the
container.
16
CA 02862545 2016-01-08
The fibre diameters were measured using a Zeiss Stemi 2000-C microscope. The
mean of 10
single measurements was determined for this purpose.
Fleece samples of 10x10 cm2 were used to determine the weight per unit area.
The weights were
determined using a micro-analytical scale made by Sartorius (model Acculab VIC-
123).
The thickness of the fleece samples was determined using a thickness measuring
device made by
Schroeder (model "Thickness gauge RAINBOW"). In this context, the
determination of the
thickness must not involve any pressure acting on the fleece to avoid any
unintended compres-
sion of the fleece and ensuing decrease of the volume.
The porosity E of the samples was calculated according to the following
formula:
PV fleece
E = 1
PBulk
mV fleece
VV fleece
=1¨
ma mb= Mc
=+
Pa = (Ma + Mb -F Mc) + Pb Pc
mb + Mc) mb mc)
Here, pfleece is the density of the non-compressed biodegradable fleece, pBuik
is the density of the
fibres 1 of the biodegradable fleece,nxece _s i the mass of the biodegradable
fleece, Vfleece is the
volume of the biodegradable fleece, pa is the density of the fibre-forming
polymer, pb is the den-
sity of the activator of secondary haemostasis, pc is the density of the
inhibitor of fibrinolysis, ma
is the mass of the fibre-forming polymer in the fleece, mb is the mass of the
activator of secon-
dary haemostasis in the fleece, and mc is the mass of the inhibitor of
fibrinolysis in the fleece. If
further components are present in the biodegradable fleece, such as, for
example, an additional
buffer substance or antibiotics, and the porosity of the fleece is to be
determined, further parame-
ters need to be taken into consideration according to the same pattern as
shown above, i.e. the
masses (ma, me, ...) and densities (pd, Pe, ...) of the additional components.
The mean pore radius
was calculated according to S. J. Eichhorn, W. W. Sampson, Statistical
geometry of pores and
statistics of porous nanofibrous assemblies, J. R. Soc. Interface, 2005; 2;
309-318. Formula 6.1
on page 315 was used for this purpose.
17
CA 02862545 2016-01-08
The fibre surface Ofibre was calculated according to the following formula:
1 ¨ V fleece
fibre = 7T = fibre 2
(Of ibre)
rr
2
whereby 0.fibre is the average diameter of the fibres 1. The contact angle was
determined using a
goniometer G40 (made by Kriiss). For this purpose, one droplet of water was
placed on the
fleece surface an wetting angle was measured after 10 s.
Average fibre diameter Ofibre: 14 5 gm
Weight per unit area: 200 g/m2
Thickness of the samples: 2 mm
Porosity: 0.919
Mean pore radius: 0.1094 mm
Total fibre surface: 462.072 mm2
Contact angle: < 30
EXEMPLARY EMBODIMENT 2:
A gelatine fleece with anti-microbial coating according to Fig. 3 was produced
by means of a
rotation spinning method as follows:
Firstly, a 24% gelatine solution was prepared. A gelatine of type A PIGSKIN
made by GELITA
AG was used. The gelatine was stirred in water. The pH was adjusted to 7.4
with NaOH (product
number: 3306576, Sigma-Aldrich, Germany). A total of 1 % by weight calcium
chloride (prod-
uct number: 102382, Merck, Germany), 1 % by weight calcium carbonate (product
number:
C4830, Sigma-Aldrich, Germany), 1 % by weight glycerol (product number: 01873,
Sigma-
Aldrich, Germany), 0.5 % by weight trans-4-aminomethylcyclohexylcarboxylic
acid (product
18
CA 02862545 2014-07-24
number: 857653, Sigma-Aldrich, Germany) and 10 mg bromocresol purple (product
number:
114375, Sigma-Aldrich, Germany) were added to the gelatine solution.
The solution was then allowed to stand uninterrupted for approximately one
hour to swell. Then,
the gelatine solution was treated at 60 C in an ultrasonic bath and then
maintained at a tempera-
ture of 80 C to 85 C for approx. 1 hour.
The gelatine solution maintained at 80 C to 85 C was guided, as the fibre
raw material, by
means of a syringe pump into the container of a device for rotation spinning
according to
DE 10 2005 048 939 A1. A second syringe pump was used concurrently to guide
polyethylene
glycol diglycidylether (product number: 475696, Sigma-Aldrich, Germany) into
the container.
The temperature of the container is approx. 120 C and the container rotates
at a speed of 4,500
rpm. Inside the container there are recesses designed to be holes with a
diameter of 0.3 mm. The
centrifugal force pressed the fibre raw material through said recesses and
spins it into fibres 1
that are drawn by means of an aspiration facility. The aspiration facility was
situated below the
container.
The fleece thus obtained was sprayed with a gentamicin palmitate solution
(Heraeus Medical,
Germany) (5 g dissolved in 100 ml methanol) and dried in a vacuum.
EXEMPLARY EMBODIMENT 3:
A gelatine fleece with anti-microbial substance and antiseptic coating
according to Fig. 4 was
produced by means of a rotation spinning method as follows:
Firstly, a 24% gelatine solution was prepared. Presently, a gelatine of type A
PIGSKIN made by
GELITA AG was used, whereby other types of gelatine can be used just as well.
The gelatine
was stirred in water. The pH was adjusted to 7.4 with NaOH (product number:
3306576, Sigma-
Aldrich, Germany). A total of 1 % by weight calcium chloride (product number:
102382, Merck,
19
. .
CA 02862545 2014-07-24
Germany), 5 % by weight calcium carbonate (product number: C4830, Sigma-
Aldrich, Ger-
many), 1 % by weight glycerol (product number: 01873, Sigma-Aldrich, Germany)
and 0.5 % by
weight 6-aminohexanoic acid (product number: 800145, Sigma-Aldrich, Germany)
and 10 mg
bromocresol purple (product number: 114413, Sigma-Aldrich, Germany) were added
to the gela-
tine solution.
The solution was then allowed to stand uninterrupted for approximately one
hour to swell. Then,
the gelatine solution was dissolved at 60 C in an ultrasonic bath and then
maintained at a tem-
perature of 80 C to 85 C for approx. 1 hour. A total of 6 % by weight
gentamicin sulfate (prod-
uct number: 345814, Merck, Germany) were then dissolved while stirring in the
hot gelatine so-
lution thus produced.
The gelatine solution maintained at 80 C to 85 C was guided, as the fibre
raw material, by
means of a syringe pump into the container of a device for rotation spinning
according to
DE 10 2005 048 939 A1.
The temperature of the container is approx. 120 C and the container rotates
at a speed of 4,500
rpm. Inside the container there are recesses designed to be holes with a
diameter of 0.3 mm. The
centrifugal force pressed the fibre raw material through said recesses and
spins it into fibres 1
that are drawn by means of an aspiration facility. The aspiration facility was
situated below the
container.
The fleece thus obtained was then stored for 12 hours at room temperature in a
desiccator in the
presence of a 36% formaldehyde solution (product number: F8775, Sigma-Aldrich,
Germany)
and then evacuated for another 72 hours to fully remove the excess of
formaldehyde.
Then, the fleece was sprayed with a polyhexanide solution (Hangzhou Dayangchem
Co., Ltd.,
China) (5 g polyhexanide in 100 ml of an ethanol/water mixture (80/20; v/v)
and dried in a vac-
uum.
Referring to further advantageous refinements and developments of the teaching
according to the
invention, reference shall be made to the general part of the description as
well as to the ap-
. =
CA 02862545 2014-07-24
pended patent claims. Finally, it shall be noted that the exemplary
embodiments have been se-
lected at random and only serve to illustrate the teaching according to the
invention without the
invention being limited to said exemplary embodiments in any way, manner or
shape.
The features of the invention disclosed in the preceding description and in
the claims, figures,
and exemplary embodiments, can be essential for the implementation of the
various embodi-
ments of the invention both alone and in any combination.
LIST OF REFERENCE NUMBERS
1 Fibre/filament
21
